Auxiliary device for water immersion lens of two-photon microscope

Abstract

The invention belongs to the technical field of two-photon microscope imaging, and specifically discloses an auxiliary device for a water immersion lens of a two-photon microscope, comprising a water replenishing bucket, a cooling interlayer and a liquid supply component; the water replenishing bucket is used to be placed on an observation window and forms a water containing cavity with the observation window, and the water containing cavity is used to place the water immersion lens of the two-photon microscope; the cooling interlayer is arranged around the outer side of the water replenishing bucket and is located at the lower half of the water replenishing bucket, and its bottom surface is located at the same horizontal plane as the bottom surface of the water replenishing bucket and forms a fitting surface, and the fitting surface is used to be in close contact with the observation window; the invention forms a water immersion lens placing container with a cooling function through the water replenishing bucket and the cooling interlayer, and cooperates with the two containers to realize continuous water replenishment of the water immersion lens, and can effectively reduce the temperature around the water immersion lens, can keep the water immersion lens always immersed in water during use, cool the lens at the same time, and prevent excessive water as immersion liquid from flowing to the periphery.

Description

Technical Field

[0001] The invention belongs to the technical field of two-photon microscope imaging, and in particular relates to an auxiliary device for a water immersion lens of a two-photon microscope. Background Art

[0002] The two-photon microscope has the advantages of low phototoxicity and high penetration of two-photons, and combined with a water immersion lens, it makes it possible to observe deeper tissue cells in living animals. Because the refractive index of light in water is closer to that of glass than in air, the water immersion objective uses water as the immersion liquid, which can expand the numerical aperture of the lens and provide a larger working distance. When using a two-photon microscope to observe specimens that are not culture dishes or perfusion tanks, especially when observing local tissues of living animals, the water immersion lens needs to use water as the medium, and the bottom of the lens needs to be immersed in the water at all times; but at the same time, because the two-photon microscope lens generates heat during use, the water around the lens evaporates quickly, and water needs to be added continuously during the observation process.

[0003] At present, most water immersion lenses of two-photon microscopes do not have special water-adding devices. Especially when observing the parts of living animals, observers usually add water by themselves with a dropper. This has the following problems: First, untimely water addition will cause the area around the lens to dry out, resulting in the water immersion lens being unable to be used normally, and observation will be interrupted until water is added again; second, if too much water is added, the water will overflow, affecting the animals on the stage or other surrounding facilities, affecting observation and other operations. Summary of the invention

[0004] In view of the above-mentioned problems, the purpose of the present invention is to provide an auxiliary device for a two-photon microscope water immersion lens, which can continuously replenish water to the water immersion lens and effectively avoid unnecessary impact on animals on the stage or other surrounding facilities after water flows, which is more conducive to subsequent observations and other operations.

[0005] The technical solution of the present invention is: an auxiliary device for a water immersion lens of a two-photon microscope, comprising:

[0006] The water supply hopper is a funnel-shaped structure, which is used to be placed on the observation window and to form a water-containing cavity with the observation window, and the water-containing cavity is used to place the water immersion lens of the two-photon microscope; a water injection port is provided on the water supply hopper;

[0007] The cooling interlayer is wound around the outer side of the water supply bucket and is located at the lower half of the water supply bucket; the cooling interlayer is provided with a liquid inlet and a liquid outlet; the bottom surface of the cooling interlayer and the bottom surface of the water supply bucket are located at the same horizontal plane and form a fitting surface, and the fitting surface is used to be in close contact with the observation window;

[0008] The liquid supply component is connected to the water injection port for providing pure water to the water containing cavity, and is respectively connected to the liquid inlet and the liquid outlet to form a circulating water circuit for providing coolant to the cooling interlayer.

[0009] Furthermore, an annular groove is provided on the bottom surface of the cooling interlayer away from the bottom of the water supply hopper, and the annular groove is used to contact the observation window by pressing and form a negative pressure area.

[0010] Furthermore, a pressing ring is provided on the bottom surface of the cooling interlayer, and the pressing ring is located directly above the annular groove.

[0011] Furthermore, the bottom surface of the cooling interlayer is provided with an inlay groove along a horizontal plane, the inlay groove is located above the annular groove, and one end of the inlay groove forms an inlay opening at a side end of the bottom surface of the cooling interlayer;

[0012] The pressing ring comprises:

[0013] A ring plate, wherein one side of the inner ring is embedded in the embedding groove, and the upper surface of the ring plate is fixed to the upper end surface of the embedding groove, and one side of the outer ring of the ring plate is exposed outside the cooling interlayer;

[0014] The pressing ring block is arranged on the outer ring of the ring plate.

[0015] Furthermore, the liquid supply assembly comprises:

[0016] A first container, wherein pure water is stored inside, and is connected to the water inlet through a peristaltic pump and a connecting hose to provide pure water to the water containing cavity;

[0017] The second container stores cooling liquid inside and is respectively connected to the liquid inlet and the liquid outlet to form a circulating water circuit for providing cooling liquid to the cooling interlayer.

[0018] Furthermore, the peristaltic pump is used to add the pure water in the first container into the water containing cavity in the form of drip irrigation.

[0019] Furthermore, the coolant is cooled pure water, and a cooling device is provided in the second container, and the cooling device is used to cool the pure water in the second container;

[0020] The overflow port is connected to the second container.

[0021] Furthermore, a water level warning line is provided on the water replenishment bucket, and an overflow port is provided on the water replenishment bucket just above the water level warning line, and the overflow port is used to discharge the pure water above the water level warning line out of the water containing cavity.

[0022] Furthermore, the water replenishment hopper and the cooling interlayer are an integrated structure.

[0023] Furthermore, the water supply hopper and cooling interlayer are both made of TPR material.

[0024] The working method of the present invention is as follows: before use, the two-photon microscope water immersion lens is placed in a water chamber, and the horizontal height of its lower end is ensured to be lower than the water level warning line, and the pure water in the first container is first added into the water chamber until the water level reaches the water level warning line.

[0025] When it starts to be used, the pure water in the first container is continuously added to the water cavity through the water injection port in the form of drip irrigation; the cooled pure water in the second container is added to the cooling interlayer through the liquid inlet, and then flows back to the second container through the liquid outlet to form a circulating water circuit. If the water level in the water cavity is higher than the water level warning line, the pure water in the water cavity flows into the second container through the overflow port for secondary use.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] The present invention forms a water immersion lens placement container with a cooling function through a water replenishing bucket and a cooling interlayer. The two containers cooperate to achieve continuous water replenishment for the water immersion lens and can effectively reduce the temperature around the water immersion lens. The water immersion lens can be kept immersed in water during use, while the lens is cooled and excessive water as immersion liquid is effectively prevented from flowing to the periphery, effectively avoiding unnecessary influence on animals on the stage or other surrounding facilities after the water flows, which is more conducive to subsequent observation and other operations. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 It is a schematic diagram of the overall structure of the present invention;

[0029] Figure 2 It is a structural schematic diagram of the water replenishment bucket and the cooling interlayer in Example 1 of the present invention;

[0030] Figure 3 It is a structural schematic diagram of the water replenishment bucket and the cooling interlayer of Example 2 of the present invention;

[0031] Figure 4 This is a partial structural exploded diagram of the water replenishment bucket and the cooling interlayer of Example 2 of the present invention;

[0032] Figure 5 It is a schematic diagram of the partial structure of the pressing ring of Example 2 of the present invention;

[0033] Figure 6 1 is a schematic diagram of the principle of a water immersion lens in an application example of the present invention using a traditional method;

[0034] Figure 7 It is a schematic diagram of the principle of the auxiliary device used in the water immersion lens of the application example of the present invention.

[0035] Among them, 1-water filling bucket, 100-card slot, 101-sealing ring, 11-water filling port, 12-water level warning line, 13-overflow port, 2-cooling interlayer, 20-ring groove, 21-liquid inlet, 22-liquid outlet, 23-pressing ring, 231-ring plate, 232-pressing ring block, 24-mounting groove, 240-mounting port, 3-first container, 4-second container, 5-observation window, 6-two-photon microscope water immersion lens, 7-dropper. DETAILED DESCRIPTION

[0036] The following is combined with Figure 1 To the attached Figure 7 , the specific embodiments of the present invention are described in detail. In the description of the present invention, it should be understood that the terms "center", "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside" and the like indicate positions or positional relationships based on the positions or positional relationships shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the present invention.

[0037] The terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features; in the description of the present invention, unless otherwise specified, "plurality" means two or more.

[0038] It should be noted that the circuit connections involved in the present invention all adopt conventional circuit connection methods and do not involve any innovation.

[0039] Example 1

[0040] like Figure 1 An auxiliary device for a water immersion lens of a two-photon microscope is shown, comprising a water supply bucket 1, a cooling interlayer 2 and a liquid supply component.

[0041] The water replenishment bucket 1 is a funnel-shaped structure, which is used to be placed on the observation window 5 and form a water-containing cavity with the observation window 5, and the water-containing cavity is used to place the water immersion lens 6 of the two-photon microscope; a water injection port 11 is provided on the water replenishment bucket 1; the cooling interlayer 2 is arranged around the outside of the water replenishment bucket 1 and is located in the lower half of the water replenishment bucket 1; a liquid inlet 21 and a liquid outlet 22 are provided on the cooling interlayer 2; the bottom surface of the cooling interlayer 2 and the bottom surface of the water replenishment bucket 1 are located in the same horizontal plane and form a fitting surface, and the fitting surface is used to be in close contact with the observation window 5; the liquid supply component is connected to the water injection port 11 to provide pure water to the water-containing cavity, and the liquid supply component is also connected to the liquid inlet 21 and the liquid outlet 22 respectively to form a circulating water circuit, which is used to provide coolant to the cooling interlayer 2. Among them, if Figure 2 As shown, the liquid inlet 21 and the liquid outlet 22 are both located on the outer wall of the cooling interlayer 2, and the liquid inlet 21 is located at the upper end of the outer wall, and the liquid outlet 22 is located at the lower end of the outer wall. It should be noted that the material of the observation window 5 is glass.

[0042] Preferably, the liquid supply assembly includes a first container 3 and a second container 4. The first container 3 stores pure water, which is connected to the water injection port 11 through a peristaltic pump and a connecting hose to provide pure water to the water cavity; the second container 4 stores coolant, which is connected to the liquid inlet 21 and the liquid outlet 22 to form a circulating water circuit, and is used to provide coolant to the cooling interlayer 2.

[0043] It should be noted that: Figure 1 As shown, this embodiment uses a commercially available double-pump-head peristaltic pump, the first container 3 is connected to the water injection port 11 through one of the pump heads, and the second container 4 is connected to the liquid inlet 21 through the other pump head.

[0044] Preferably, the peristaltic pump is used to drip the pure water in the first container 3 into the water containing cavity. The peristaltic pump is used to drip water into the water containing cavity in a continuous and slow manner, thereby ensuring that the water immersion lens is always submerged in water during use.

[0045] Preferably, the coolant is cooled pure water, and a cooling device is provided in the second container 4, and the cooling device is used to cool the pure water in the second container 4; the overflow port 13 is connected to the second container 4. The water flowing out of the overflow port 13 is added to the cold water cycle for use, so as to achieve secondary utilization and avoid waste. The cooling device adopts a commercially available cooling device.

[0046] Preferably, in order to effectively prevent excessive water from overflowing from the water holding chamber and causing water to overflow and affect the animals on the loading platform or other surrounding facilities, a water level warning line 12 is also provided on the water filling bucket 1, and an overflow port 13 is provided on the water filling bucket 1 just above the water level warning line 12. The overflow port 13 is used to discharge pure water above the water level warning line 12 out of the water holding chamber.

[0047] Preferably, the water supply hopper 1 and the cooling interlayer 2 are an integrated structure, and the integrated structure makes the formed fitting surface integrated, which can effectively fit with the observation window 5, thereby avoiding the problem of a gap between the two resulting in cooling liquid leakage caused by a non-integrated structure.

[0048] Preferably, the water supply hopper 1 and the cooling interlayer 2 are both made of TPR material. The TPR material itself has plasticity and surface adhesion, so that the bonding surface can be automatically and effectively bonded to the glass observation window 5 after being placed on the observation window 5, which has the advantage of preventing water from overflowing.

[0049] Example 2

[0050] The difference from Example 1 is:

[0051] Preferably, Figure 3 , Figure 4 As shown, the bottom surface of the cooling interlayer 2 is provided with an annular groove 20 away from the bottom of the water supply hopper 1. The annular groove 20 is used to contact the observation window 5 by pressing and form a negative pressure area. Based on the suction cup structure principle, the annular groove 20 is pressed to form a vacuum cavity above the observation window 5, so that the bonding surface can be well adsorbed on the observation window 5 to achieve the effect of preventing water from overflowing.

[0052] Preferably, Figure 3 , Figure 4 As shown, a pressing ring 23 is provided on the bottom surface of the cooling interlayer 2, and the pressing ring 23 is located directly above the annular groove 20. In actual operation, the annular groove 20 can be squeezed by only pressing the pressing ring 23, which is more convenient to operate.

[0053] Preferably, Figure 3 , Figure 4 , Figure 5 As shown, the bottom surface of the cooling interlayer 2 is provided with an inlay groove 24 along a horizontal plane, the inlay groove 24 is located above the ring groove 20, and one end of the inlay groove 24 forms an inlay opening 240 at the side end of the bottom surface of the cooling interlayer 2; the pressing ring 23 includes a ring plate 231 and a pressing ring block 232. The inner ring side of the ring plate 231 passes through the inlay opening 240 and is inlaid into the inlay groove 24, and the upper surface of the ring plate 231 is fixed to the upper end surface of the inlay groove 24, and the outer ring side of the ring plate 231 is exposed outside the cooling interlayer 2; the pressing ring block 232 is arranged on the outer ring of the ring plate 231.

[0054] In actual use, the ring plate 231 is set on the upper end surface of the inlay groove 24, and the lower end surface of the inlay groove 24 is not connected to the ring plate 231. When the pressing ring block 232 is pressed, the ring plate 231 contacts and squeezes the lower end surface of the inlay groove 24, causing the ring groove 20 to deform and adsorb on the observation window 5. After the pressing of the pressing ring block 232 is stopped, since the ring plate 231 is not connected to the lower end surface of the inlay groove 24, the reset process of the ring plate 231 does not drive the lower end surface of the inlay groove 24 to reset.

[0055] Application Examples

[0056] An experiment to observe the mouse brain using a water immersion lens.

[0057] like Figure 6 As shown, water is added between the water immersion lens and the observation window 5 in a traditional way using a dropper 7. At this time, due to excessive water addition, water overflows, seriously affecting the animals on the stage.

[0058] like Figure 7 As shown, the auxiliary device proposed in Example 1 is used to add water between the water immersion lens and the observation window 5. Before use, the fitting surface is closely contacted with the observation window 5. Then the two-photon microscope water immersion lens is placed in the water chamber, and the horizontal height of its lower end is ensured to be lower than the water level warning line 12. First, pure water in the first container 3 is added into the water chamber until the water level reaches the water level warning line 12.

[0059] When it starts to be used, the pure water in the first container 3 is continuously added to the water chamber through the water injection port 11 in the form of drip irrigation; and the cooled pure water in the second container 4 is added to the cooling interlayer 2 through the liquid inlet 21, and then flows back to the second container 4 through the liquid outlet 22 to form a circulating water circuit. If the water level in the water chamber is higher than the water level warning line 12, the pure water in the water chamber flows into the second container 4 through the overflow port 12 for secondary use. This effectively ensures the continuous replenishment of the water immersion lens, and can effectively reduce the temperature around the water immersion lens, so that the water immersion lens can be kept immersed in water during use, while cooling the lens and preventing too much water as immersion liquid from flowing to the surrounding area.

[0060] The specific models of the above electronic components are not specially specified, and ordinary products available on the market can be selected as long as they can meet the use requirements of the present invention.

[0061] The specific embodiments described above further illustrate the objectives, technical solutions and beneficial effects of the present invention in detail. It should be understood that the above description is only a specific embodiment of the present invention and does not limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention are included in the protection scope of the present invention.

Claims

1. An auxiliary device for a two-photon microscope water immersion lens, characterized in that: include: The water supply hopper (1) is a funnel-shaped structure. The water supply hopper (1) is used to be placed on the observation window (5) and to form a water-containing cavity with the observation window (5). The water-containing cavity is used to place a two-photon microscope water immersion lens (6). A water injection port (11) is provided on the water supply hopper (1). The cooling interlayer (2) is arranged around the outside of the water supply bucket (1) and is located at the lower half of the water supply bucket (1); the cooling interlayer (2) is provided with a liquid inlet (21) and a liquid outlet (22); the bottom surface of the cooling interlayer (2) and the bottom surface of the water supply bucket (1) are located at the same horizontal plane and form a fitting surface, and the fitting surface is used to be in close contact with the observation window (5); A liquid supply component, connected to the water injection port (11) for supplying pure water into the water containing cavity, and connected to the liquid inlet (21) and the liquid outlet (22) respectively to form a circulating water circuit for supplying cooling liquid into the cooling interlayer (2); An annular groove (20) is provided on the bottom surface of the cooling interlayer (2) away from the bottom of the water supply bucket (1), and the annular groove (20) is used to contact the observation window (5) by pressing and form a negative pressure area; The liquid supply assembly comprises: A first container (3) stores pure water therein and is connected to the water injection port (11) via a peristaltic pump and a connecting hose for providing pure water to the water containing cavity; The second container (4) stores cooling liquid therein and is respectively connected to the liquid inlet (21) and the liquid outlet (22) to form a circulating water circuit for providing cooling liquid to the cooling interlayer (2).

2. The auxiliary device for a two-photon microscope water immersion lens according to claim 1, characterized in that: A pressing ring (23) is provided on the bottom surface of the cooling interlayer (2), and the pressing ring (23) is located directly above the annular groove (20).

3. The auxiliary device for a two-photon microscope water immersion lens according to claim 2, characterized in that: The bottom surface of the cooling interlayer (2) is provided with an inlay groove (24) along a horizontal plane, the inlay groove (24) is located above the annular groove (20), and one end of the inlay groove forms an inlay opening (240) at a side end of the bottom surface of the cooling interlayer (2); The pressing ring (23) comprises: An annular plate (231), wherein one side of the inner ring is embedded in the embedding groove (24), and the upper surface of the annular plate (231) is fixed to the upper end surface of the embedding groove (24), and one side of the outer ring of the annular plate (231) is exposed outside the cooling interlayer (2); The pressing ring block (232) is arranged on the outer ring of the ring plate (231).

4. The auxiliary device for a two-photon microscope water immersion lens according to claim 1, characterized in that: The peristaltic pump is used to add the pure water in the first container (3) into the water containing cavity in the form of drip irrigation.

5. The auxiliary device for a two-photon microscope water immersion lens according to claim 1, characterized in that: The coolant is cooled pure water, and a cooling device is provided in the second container (4), and the cooling device is used to cool the pure water in the second container (4); The overflow port (13) is connected to the second container (4).

6. The auxiliary device for a two-photon microscope water immersion lens according to claim 1, characterized in that: The water supply hopper (1) is also provided with a water level warning line (12), and the water supply hopper (1) is provided with an overflow port (13) directly above the water level warning line (12), and the overflow port (13) is used to discharge the pure water above the water level warning line (12) out of the water containing cavity.

7. The auxiliary device for a two-photon microscope water immersion lens according to claim 1, characterized in that: The water supply hopper (1) and the cooling interlayer (2) are an integrated structure.

8. The auxiliary device for a two-photon microscope water immersion lens according to claim 1, characterized in that: The water supply hopper (1) and the cooling interlayer (2) are both made of TPR material.

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